Method for preparing cosmetic raw material with anti-inflammatory, soothing and repairing effects through microbial fermentation method

Abstract

The invention belongs to the technical field of cosmetic raw materials, and particularly discloses a method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects through a microbial fermentation method, and the method comprises the following steps: S1, preparing a fermentation culture medium; s2, preparing fermentation liquor: culturing saccharomyces cerevisiae and / or lactobacillus plantarum, and then inoculating the saccharomyces cerevisiae and / or lactobacillus plantarum into the fermentation culture medium obtained in the step S1 to obtain the fermentation liquor; s3, sieving the fermentation liquor obtained in the step S2 to obtain an upper-layer active raw material and a lower-layer substance; centrifuging the obtained lower-layer substance, and performing freeze drying treatment to obtain a lower-layer active raw material; and S4, mixing the upper-layer active raw material and the lower-layer active raw material obtained in the step S3, and then adding calculus bovis factitius. According to the invention, a specific strain is utilized to ferment a specific fermentation culture medium raw material, and the obtained cosmetic raw material active ingredients have very good anti-inflammatory, soothing and repairing effects; and the preparation process is simple and easy for large-scale production.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and specifically relates to a method for preparing cosmetic raw materials with anti-inflammatory, soothing and repairing effects by a microbial fermentation method. Background Art

[0002] With the growth of market demand, consumers' expectations for cosmetics are no longer limited to a single function, but have turned to the hope of using compound products with multiple functions. Based on this, some cosmetic manufacturers on the market have adopted active ingredients with different functions obtained by microbial fermentation, and then mixed them to prepare finished cosmetics with multiple functions. Microbial fermentation is a process of converting natural raw materials into highly active ingredients through the metabolic activities of microorganisms (such as bacteria, fungi, yeast, etc.). This technology not only improves the efficacy and safety of ingredients, but also promotes the development of green skin care.

[0003] In the prior art, for example, Chinese patent document CN118526434A discloses a whitening fermented puree prepared by fermenting seven Chinese herbal medicines such as pomegranate peel, peony root, and salvia miltiorrhiza with Bifidobacterium bifidum. The fermented puree can significantly improve the antioxidant capacity of the Chinese herbal medicine composition by scavenging DPPH free radicals, and can also inhibit tyrosinase activity to improve the whitening effect.

[0004] For example, Chinese patent document CN 118620982A relates to a preparation method and application of a Panax notoginseng root fermentation liquid with increased ginsenoside Rh1 content. The Panax notoginseng root is subjected to alcohol extraction and ultrasound to obtain a crude sample extract, which is then subjected to column chromatography to obtain an enrichment liquid. A strain is inoculated into the enrichment liquid for fermentation to obtain a Panax notoginseng root fermentation liquid with increased ginsenoside Rh1 content. The fermentation liquid can be used as a cosmetic raw material with antioxidant, anti-wrinkle and firming effects.

[0005] For example, Chinese patent document CN118662400A provides an application of a bifidobacterium longum fermentation product lysate in repairing, regulating skin barrier and soothing and anti-inflammatory. The bifidobacterium longum fermentation lysate can improve the three-layer barrier function of the skin: microbial barrier, physical barrier and immune barrier, thereby achieving the purpose of relieving skin inflammation, repairing and regulating skin barrier damage.

[0006] From the above literature, we can see that microbial fermentation based on common substances has been reported; how to use specific substances for microbial fermentation to obtain good cosmetic effects is still worthy of continuous exploration and research.

[0007] Based on this, the present invention is proposed. Summary of the invention

[0008] In view of the shortcomings of the prior art, one of the purposes of the present invention is to provide a method for preparing cosmetic raw materials with anti-inflammatory, soothing and repairing effects by microbial fermentation, which mainly uses fermentation technology and has a simple and practical preparation process.

[0009] The second object of the present invention is to provide a cosmetic raw material prepared by the above method, which is prepared from the traditional Mayinglong hemorrhoid ointment classic raw materials and has safe and common ingredients.

[0010] The third object of the present invention is to provide the use of the above-mentioned cosmetic raw materials in the preparation of cosmetics, relying on the anti-inflammatory, soothing and repairing effects of the cosmetic raw materials to give the prepared cosmetics corresponding effects.

[0011] In order to achieve the above object, the present invention adopts the following technical solution:

[0012] In a first aspect, the present invention provides a method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0013] S1. Preparation of fermentation medium;

[0014] S2, preparation of fermentation broth: culturing Saccharomyces cerevisiae and / or Lactobacillus plantarum, and then inoculating into the fermentation medium obtained in step S1 to obtain a fermentation broth;

[0015] S3, sieving the fermentation liquid obtained in step S2 to obtain an upper layer of active raw materials and a lower layer; centrifuging the obtained lower layer and freeze-drying it to obtain a lower layer of active raw materials;

[0016] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3, and then adding artificial bezoar.

[0017] In the above technical solution, specific raw materials are first made into fermentation medium, and then added with specific bacterial flora to obtain fermentation liquid, and then screened, separated and mixed to obtain the raw materials of cosmetic active ingredients. Among them, the enzymes produced during the growth of Saccharomyces cerevisiae and Lactobacillus plantarum can promote specific metabolic pathways and produce a variety of active substances with specific effects, such as glutathione (GSH) and γ-aminobutyric acid (GABA), thereby improving the anti-inflammatory, soothing and repairing effects of cosmetics.

[0018] More preferably, in step S1, the preparation step comprises: mixing basic zinc hydroxide, pearl powder, and amber powder after drying to obtain a mixture; then crushing the mixture, adding a carbon source, a nitrogen source, an inorganic salt, and water, and sterilizing to obtain a fermentation medium. In the above-mentioned fermentation medium, the basic zinc hydroxide, pearl powder, and amber powder used are classic components of the hemorrhoid cream of Mayinglong Pharmaceutical Group. There is no report on their application in microbial flora fermentation. Therefore, this is the first creation of the applicant and has achieved very good cosmetic use effects.

[0019] More preferably, the mass proportion of basic zinc hydroxide in the mixture is 95-98.5%, the mass proportion of pearl powder is 1.2-3.5%, and the mass proportion of amber powder is 0.3-1.5%;

[0020] In the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder and amber powder is 5 to 50 g / L.

[0021] More preferably, the added amounts of the carbon source, nitrogen source and inorganic salt are 15-25 g / L, 15-25 g / L and 0.5-5 g / L, respectively.

[0022] More preferably, the carbon source is selected from one of glucose, sucrose, maltose, starch, and galactose; the nitrogen source is selected from one of soy peptone, peanut meal, and cottonseed protein; and the inorganic salt is selected from one of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, and ammonium dihydrogen phosphate.

[0023] More preferably, in step S2, the Saccharomyces cerevisiae is cultured in a modified YPD medium (whose main components are: 20 g / L glucose and 20 g / L soy peptone, which is named YSP medium), cultured at 25-30° C. for 16-20 h, and then inoculated into the fermentation medium at an inoculum amount of 10-15%;

[0024] Lactobacillus plantarum was cultured using a modified MRS medium (mainly composed of 20 g / L glucose, 20 g / L soy peptone, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium citrate, 5 g / L sodium acetate, 0.58 g / L magnesium sulfate, 0.25 g / L manganese sulfate, and 1 mL Tween 80, named MSPR medium), cultured at 35-40° C. for 16-20 h, and then inoculated into a fermentation medium at an inoculum size of 10-15%.

[0025] For example, the YSP culture medium mentioned above can be purchased commercially or homemade, and the MSPR culture medium can also be purchased commercially or homemade.

[0026] In some embodiments, the fermentation of step S2 is preferably carried out using both Saccharomyces cerevisiae and Lactobacillus plantarum. The early stage of the fermentation culture of Saccharomyces cerevisiae is mainly the accumulation of yeast biomass, and the secondary metabolite glutathione is synthesized in the later stage (48-96h). Glutathione has strong antioxidant properties and can remove free radicals and reactive oxygen species in the body, thereby improving skin inflammation problems caused by ultraviolet rays. The early stage of the fermentation culture of Lactobacillus plantarum is mainly the accumulation of lactic acid bacteria biomass, and the secondary metabolite γ-aminobutyric acid is synthesized in the later stage (48-96h). γ-Aminobutyric acid (GABA) reduces the release of inflammatory factors such as tumor necrosis factor α (TNF-α), interleukin 1β (1L-1β) and interleukin 6 (1L-6) by inhibiting the NF-kB signaling pathway to reduce the release of inflammatory factors such as tumor necrosis factor α (TNF-α), interleukin 1β (1L-1β) and interleukin 6 (1L-6) to reduce skin inflammatory reactions; γ-aminobutyric acid can also promote skin fibroblasts to synthesize collagen and hyaluronic acid, which helps to repair and maintain the integrity of the skin barrier.

[0027] More preferably, in step S3, the sieve mesh is 350-450 mesh, preferably 400 mesh; the centrifugal speed is 6000-10000 rpm, and the centrifugal time is 3-10 min; preferably, the centrifugal speed is 8000 rpm, and the centrifugal time is 5 min. In step S3, the fermentation liquid with cells is filtered out, and the unfermented cosmetic initial raw materials and solid substances that may be produced during the fermentation process are retained; centrifugation is used to remove cells, reduce the effects of nucleic acids and other substances in cells on the skin state, and maintain the normal state of the skin.

[0028] More preferably, in step S4, the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 10-90:10-40:5-20.

[0029] In some embodiments, the mixing in step S4 can be performed in the following ways including but not limited to: three-dimensional mixing, double cone mixing, ball milling mixing, etc.

[0030] In a second aspect, the present invention provides a cosmetic raw material prepared by the above method.

[0031] In a third aspect, the present invention provides the use of the above-mentioned cosmetic raw materials in the preparation of cosmetics. For example, the fermentation product upper layer active raw materials, the fermentation product lower layer active raw materials and artificial bezoar in the present invention can be mixed in a specific ratio and added to the cream as an active ingredient, aiming to improve the anti-inflammatory, soothing and repairing effects of cosmetics and better meet consumers' demand for efficient skin care.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] (1) The method for preparing cosmetic raw materials provided by the present invention creatively uses basic zinc hydroxide, pearl powder and amber powder as raw materials for fermentation medium, and then selects brewer's yeast and lactobacillus plantarum for co-fermentation, thereby improving the dissolution and conversion of effective ingredients, improving the bioavailability of raw materials, and producing a variety of active substances with specific functions, such as glutathione (GSH) and gamma-aminobutyric acid (GABA). The obtained fermentation product has good anti-inflammatory, soothing and repairing effects.

[0034] (2) In the present invention, the fermentation product is mixed with artificial bezoar to prepare a cosmetic composition. The glutathione in the fermentation product reduces oxidative stress, the γ-aminobutyric acid calms the skin, and the artificial bezoar inhibits inflammatory response. The three form a variety of protective networks, accelerate the recovery of the skin barrier, and further enhance the anti-inflammatory, soothing and repairing effects of the cosmetic composition.

[0035] (3) The preparation method of the cosmetic raw material provided by the present invention is simple and efficient, and is suitable for industrial large-scale production and application.

[0036] In conclusion, the present invention utilizes specific bacterial species to ferment specific fermentation medium raw materials, and the obtained cosmetic raw material active ingredients have very good anti-inflammatory, soothing and repairing effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The figure is a schematic diagram of the preparation process of the cosmetic raw material of the present invention. DETAILED DESCRIPTION

[0038] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with embodiments. Of course, the specific embodiments described here are only used to explain the present invention and are not used to limit the present invention.

[0039] Although the steps in the present invention are arranged with numbers, they are not used to limit the order of the steps. Unless the order of the steps is clearly stated or the execution of a certain step requires other steps as a basis, the relative order of the steps can be adjusted. It is understood that the term "and / or" used in this article involves and covers any and all possible combinations of one or more of the associated listed items.

[0040] Unless otherwise specified, the raw materials in the present invention are purchased from the market, wherein the preservation number of Saccharomyces cerevisiae is CCTCC M2012215 and the preservation number of Lactobacillus plantarum is CGMCC 1.12732, both of which are purchased from the market.

[0041] Example 1

[0042] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0043] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0044] The mass proportion of basic zinc hydroxide in the mixture is 96%, the mass proportion of pearl powder is 3%, and the mass proportion of amber powder is 1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 10g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0045] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0046] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 8000 rpm for 8 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0047] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:6.

[0048] Example 2

[0049] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0050] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0051] The mass proportion of basic zinc hydroxide in the mixture is 96%, the mass proportion of pearl powder is 3%, and the mass proportion of amber powder is 1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 15g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0052] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0053] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 9000 rpm for 5 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0054] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:6.

[0055] Example 3

[0056] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0057] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0058] The mass proportion of basic zinc hydroxide in the mixture is 96%, the mass proportion of pearl powder is 3%, and the mass proportion of amber powder is 1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 20g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0059] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0060] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 8000 rpm for 5 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0061] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:5.5.

[0062] Example 4

[0063] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0064] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0065] The mass proportion of basic zinc hydroxide in the mixture is 96%, the mass proportion of pearl powder is 3%, and the mass proportion of amber powder is 1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 25g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0066] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0067] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 8000 rpm for 5 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0068] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:5.5.

[0069] Example 5

[0070] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0071] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0072] The mass proportion of basic zinc hydroxide in the mixture is 96%, the mass proportion of pearl powder is 3%, and the mass proportion of amber powder is 1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 30g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0073] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0074] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 8000 rpm for 5 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0075] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:6.5.

[0076] Example 6

[0077] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0078] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0079] The mass proportion of basic zinc hydroxide in the mixture is 96%, the mass proportion of pearl powder is 3%, and the mass proportion of amber powder is 1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 35g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0080] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0081] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 8000 rpm for 5 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0082] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:5.

[0083] Example 7

[0084] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0085] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0086] The mass proportion of basic zinc hydroxide in the mixture is 96%, the mass proportion of pearl powder is 3%, and the mass proportion of amber powder is 1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 40g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0087] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0088] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 8000 rpm for 5 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0089] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:6.

[0090] Example 8

[0091] A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, comprising the following steps:

[0092] S1. Preparation of fermentation medium: basic zinc hydroxide, pearl powder and amber powder are dried and mixed to obtain a mixture; the mixture is then crushed to 300 meshes, and then glucose, soy peptone, dipotassium hydrogen phosphate and water are added, and the mixture is sterilized in an autoclave to obtain a fermentation medium;

[0093] Among them, the mass proportion of basic zinc hydroxide in the mixture is 95.5%, the mass proportion of pearl powder is 3.4%, and the mass proportion of amber powder is 1.1%; in the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder, and amber powder is 10g / L; in the fermentation medium: glucose is 20g / L, soy peptone is 20g / L, and potassium dihydrogen phosphate is 1g / L;

[0094] S2, preparation of fermentation liquid: culturing Saccharomyces cerevisiae with YSP culture medium at 30°C for 24h, then inoculating 10% of the culture medium into the fermentation medium in step S1 and culturing for 4d to obtain a first fermentation liquid; culturing Lactobacillus plantarum with MSPR culture medium at 37°C for 24h, then inoculating 10% of the culture medium into the fermentation medium and culturing for 4d to obtain a second fermentation liquid; combining the first fermentation liquid and the second fermentation liquid to obtain a fermentation liquid;

[0095] S3, sieving the fermentation liquid obtained in step S2 through 400 mesh to separate the upper layer of insoluble matter from the fermentation liquid to obtain the upper layer of active raw materials and the lower layer; centrifuging the obtained lower layer at 8000 rpm for 8 min, taking the supernatant and freeze-drying it in a vacuum freeze dryer to obtain the lower layer of active raw materials;

[0096] S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3 and then adding artificial bezoar; wherein the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 1:9:5.5.

[0097] Comparative Example 1

[0098] Compared with Example 1, step S2 of Comparative Example 1 only includes the first fermentation liquid obtained by fermentation of Saccharomyces cerevisiae, and does not include the second fermentation liquid obtained by fermentation of Lactobacillus plantarum, and the rest are the same.

[0099] Comparative Example 2

[0100] Compared with Example 1, step S2 of Comparative Example 2 only includes the second fermentation liquid obtained by fermentation of Lactobacillus plantarum, and does not include the first fermentation liquid obtained by fermentation of Saccharomyces cerevisiae, and the rest are the same.

[0101] Comparative Example 3

[0102] Compared with Example 1, in step S3 of Comparative Example 4, the obtained lower layer is used to intercept the brewer's yeast and Lactobacillus plantarum in the fermentation broth using a small-pore nanomembrane (0.1-0.2 μm), and the obtained clear liquid is freeze-dried by a vacuum freeze dryer to obtain the lower layer active raw material, and the rest are the same.

[0103] Comparative Example 4

[0104] Compared with Example 1, in step S4 of Comparative Example 4, no artificial bezoar is added, and the rest is the same.

[0105] Test example

[0106] The cosmetic raw materials obtained in Examples 1 to 7 and Comparative Examples 1 to 4 were subjected to GABA content determination, GSH content determination and hyaluronidase activity inhibition rate determination. The GABA content in the fermentation broth was determined by Berthelot colorimetry.

[0107] Drawing of GABA standard curve: Prepare a series of GABA solutions with different concentrations, add 0.2 mL of pH 9.0, 0.2 mol / L boric acid buffer to 0.5 mL of sample, mix well, add 1 mL of 6% phenol solution and 0.5 mL of 7% NaClO solution, shake thoroughly, then place in a hot water bath for 7 min, quickly cool in an ice water bath for 9 min, add 2 mL of 60% ethanol solution, measure the absorbance at 630 nm, and make a standard curve. According to the sample absorbance measured by Berthelot colorimetry, substitute it into the standard curve to calculate the GABA content.

[0108] The GSH content in the fermentation broth was detected by high performance liquid chromatography, and the chromatographic conditions were as follows: chromatographic column: C18; mobile phase A: methanol, mobile phase B: sodium heptane sulfonate-potassium dihydrogen phosphate buffer (A:B=1:9); flow rate: 0.8 mL / min; column temperature: 30°C; detector: UV-visible detector; wavelength: 210 nm.

[0109] Drawing of GSH standard curve: Weigh 50 mg of glutathione standard, dilute to 100 mL with UP water to obtain 500 mg / L glutathione mother solution, dilute to prepare 50, 100, 200, 300, 400, 500 mg / L standard solution, detect the peak area of ​​glutathione solution with different concentrations by HPLC, draw glutathione standard curve with glutathione concentration as the horizontal axis and the peak area detected by liquid phase as the vertical axis, and obtain the calculation formula. According to the concentration of GSH in the fermentation broth measured by HPLC, the GSH content is calculated according to formula (1):

[0110] Glutathione content (mg / g) = A×V×n / M(1)

[0111] In formula (1), A is the concentration of GSH; V is the volume of bacterial suspension; n is the dilution factor; M is the dry weight of cells.

[0112] Hyaluronidase inhibition test method: Take 50 μL of sample and hyaluronidase solution (500 U / mL), shake thoroughly, and place in a 37°C water bath for 20 min. Add 10 μL of 2.5 mol / L CaCl 2The solution was shaken in a 37°C water bath for 20 min, and then 50 μL sodium hyaluronate (0.5 mg / mL) was added, and the mixture was incubated in a 37°C water bath for 40 min and at room temperature for 10 min. 50 μL distilled water, 10 μL 5 mol / L NaOH solution, and 50 μL acetylacetone solution were added, and the mixture was in a boiling water bath for 15 min, cooled for 5 min, cooled in a cold water bath for 10 min, and at room temperature for 10 min. 100 μL P-DAB color developer was added, and after sufficient shaking, 350 μL anhydrous ethanol was added to develop the color at room temperature for 30 min, and the OD530 was measured as the absorbance of tube A. Sodium hyaluronate was replaced by acetate buffer (0.1 mol / L, pH 5.4), and the OD530 was measured as the absorbance of tube B. Water was used to replace the sample, and the acetate buffer was used to replace the hyaluronidase solution, and the OD530 was measured as the absorbance of tube C. Water was used to replace the sample, and the acetate buffer was used to replace the hyaluronidase solution and sodium hyaluronate, and the OD530 was measured as the absorbance of tube D.

[0113] Hyaluronidase inhibition rate (%) = [1-(AB) / (CD)]*100% (2)

[0114] In formula (2): A-absorbance of sample solution (with sample, enzyme, sodium hyaluronate); B-absorbance of sample blank control (with acetic acid buffer instead of sodium hyaluronate); C-absorbance of control solution (with water instead of sample, and acetic acid instead of enzyme); D-absorbance of control blank solution (with water instead of sample, and acetic acid buffer instead of hyaluronidase solution and sodium hyaluronate).

[0115] The test results in the embodiments and comparative examples are shown in Table 1.

[0116] Table 1 Test results

[0117]

[0118] In Example 1, brewer's yeast and Lactobacillus plantarum were used to co-ferment the components of the ancient Babao prescription, and the GABA content was 2.03 mg / mL, the GSH content was 21.15 mg / g, and the hyaluronidase activity inhibition rate was 74.48%. Compared with Comparative Example 1, which used only brewer's yeast to ferment the components of the ancient Babao prescription, the GABA content in the fermentation liquid of Example 1 was better than that in Comparative Example 1; compared with Comparative Example 2, which used only Lactobacillus plantarum to ferment the components of the ancient Babao prescription, the GSH content in the fermentation liquid of Example 1 was better than that in Comparative Example 2; and compared with Comparative Example 1 and Comparative Example 2, the hyaluronidase activity inhibition rate of Example 1 was better than that of Comparative Example 1 and Comparative Example 2, indicating that the co-fermentation of brewer's yeast and Lactobacillus plantarum is helpful to synthesize GABA and GSH and improve the hyaluronidase activity inhibition rate.

[0119] In Example 2, Example 3, Example 4 and Example 5, compared with Example 1, as the content of the components of the ancient Babao recipe in the fermentation medium increased, the GABA content and the GSH content decreased, and the hyaluronidase activity inhibition rate increased, indicating that within a certain range, increasing the content of the components of the ancient Babao recipe can promote the hyaluronidase activity inhibition rate.

[0120] In Examples 6 and 7, compared with Example 1, the GABA content, GSH content and hyaluronidase activity inhibition rate were lower than those in Example 1, the GABA content was 1.76 mg / mL and 1.56 mg / mL, the GSH content was 17.45 mg / g and 18.45 mg / g, respectively, and the hyaluronidase activity inhibition rate was only 69.54% and 67.25%.

[0121] In Comparative Example 3, centrifugation was not used to remove cells, that is, the yeast and lactic acid bacteria produced during the fermentation process were retained. The GSH content of the cosmetic raw material obtained was 18.46 mg / g, and the GABA content and hyaluronidase activity inhibition rate were lower than those in Example 1.

[0122] In Comparative Example 4, no artificial bezoar was added, that is, the cosmetic raw materials only contained cosmetic raw material 1 and cosmetic raw material 2 (corresponding to the upper layer active raw material and the lower layer active raw material mentioned above, respectively), and the GABA content, GSH content, and hyaluronidase activity inhibition rate were lower than those in Example 1.

[0123] The results show that the new cosmetic raw material obtained by the method of this patent can act as a hyaluronidase inhibitor with a high inhibition rate, and can be used as a new cosmetic raw material with anti-inflammatory, soothing and repairing effects.

[0124] The present invention illustrates the technical concept of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of individual raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A method for preparing a cosmetic raw material with anti-inflammatory, soothing and repairing effects by a microbial fermentation method, characterized in that: The following steps are involved: S1. Preparation of fermentation medium; S2, preparation of fermentation broth: culturing Saccharomyces cerevisiae and / or Lactobacillus plantarum, and then inoculating into the fermentation medium obtained in step S1 to obtain a fermentation broth; S3, sieving the fermentation liquid obtained in step S2 to obtain an upper layer of active raw materials and a lower layer; centrifuging the obtained lower layer and freeze-drying it to obtain a lower layer of active raw materials; S4, mixing the upper layer active raw material and the lower layer active raw material obtained in step S3, and then adding artificial bezoar.

2. The method according to claim 1, characterized in that In step S1, the preparation steps include: drying basic zinc hydroxide, pearl powder, and amber powder respectively and then mixing them to obtain a mixture; then crushing the mixture, and then adding a carbon source, a nitrogen source, an inorganic salt and water, and sterilizing the mixture to obtain a fermentation medium.

3. The method according to claim 2, characterized in that The mass proportion of basic zinc hydroxide in the mixture is 95-98.5%, the mass proportion of pearl powder is 1.2-3.5%, and the mass proportion of amber powder is 0.3-1.5%; In the fermentation medium, the total concentration of basic zinc hydroxide, pearl powder and amber powder is 5 to 50 g / L.

4. The method according to claim 2 or 3, characterized in that: The added amounts of carbon source, nitrogen source and inorganic salt are 15-25 g / L, 15-25 g / L and 0.5-5 g / L respectively.

5. The method according to claim 4, characterized in that The carbon source is selected from one of glucose, sucrose, maltose, starch and galactose; the nitrogen source is selected from one of soy peptone, peanut meal and cottonseed protein; and the inorganic salt is selected from one of potassium dihydrogen phosphate, dipotassium hydrogen phosphate and ammonium dihydrogen phosphate.

6. The method according to claim 1, characterized in that In step S2, the brewer's yeast is cultured using a YSP medium at 25-30° C. for 16-20 hours, and then inoculated into a fermentation medium at an inoculum amount of 10-15%; the YSP medium comprises the following components: 20 g / L glucose and 20 g / L soy peptone; Lactobacillus plantarum is cultured in MSPR medium at 35-40° C. for 16-20 hours, and then inoculated into a fermentation medium at an inoculum amount of 10-15%; the MSPR medium includes the following components: 20 g / L glucose, 20 g / L soy peptone, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium citrate, 5 g / L sodium acetate, 0.58 g / L magnesium sulfate, 0.25 g / L manganese sulfate, and 1 mL Tween 80.

7. The method according to claim 1, characterized in that In step S3, the sieving mesh number is 350-450 meshes; the centrifugal speed is 6000-10000 rpm, and the centrifugal time is 3-10 min.

8. The method according to claim 1, characterized in that In step S4, the mass ratio of the upper layer active raw material, the lower layer active raw material and bezoar is 10-90:10-40:5-20.

9. A cosmetic raw material prepared by the method according to any one of claims 1 to 8.

10. Use of the cosmetic raw material according to claim 9 in the preparation of cosmetics.

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